Fig. 4. Gelatin zymographic analysis of culture medium. Activators and inhibitors were added directly at the beginning of culture. Medium aliqouts were collected after a culture period of 20 hours and then subjected to SDS-PAGE under nonreducing conditions on a 4.5-15% gradient gel containing 1% gelatin. The zymograph was developed and stained as described in Materials and Methods. (A) Lanes 1-4: medium of monocultured macrophages; lanes 5-7: medium of cocultures with chondrocytes and macrophages; lanes 8-9: medium of monocultured chondrocytes. Plasmin and/or aprotinin was added as indicated. (B) Lanes 1-3: coculture medium. Plasmin or plasminogen was added as indicated. (C) Lanes 1-5: coculture medium. MMP-3 inhibitor II was added as indicated.
